HUMAN MICROTUBULE-ASSOCIATED PROTEIN-2C LOCALIZES TO DENDRITES AND AXONS IN FETAL SPINAL MOTOR-NEURONS

Microtubule-associated protein-2 (MAP-2) functions to maintain neurona l morphology by promoting the assembly of microtubules. MAP-2c is an a lternately spliced form of MAP-2, containing the first 151 amino acids of high-molecular-weight (HMW) MAP-2 joined to the last 321 amino aci ds, eliminating 1,352 amino acids specific to HMW MAP-2. A polyclonal antibody generated to the splice site of human MAP-2c was used to dete rmine its cellular localization. The MAP-2c antiserum was depleted of any HMW MAP-2 reactivity by absorption with HMW MAP-2 fusion protein. Western blot analysis of human fetal spinal cord homogenates demonstra ted that the antibody is specific for human MAP-2c. MAP-2c immunoreact ivity was found in the perinuclear cytoplasm and processes of anterior motor neurons and large processes of the posterior column in sections from 22-24-week human fetal spinal cord. Double-label confocal micros copy was performed using the MAP-2c polyclonal antibody and either a H MW MAP-2 or a neurofilament protein (highly phosphorylated 160- and 20 0-kDa protein) monoclonal antibody to identify these processes as dend rites or axons, respectively. HMW MAP-2 and MAP-2c colocalized in cell bodies and dendrites of anterior motor neurons, demonstrating for the first time the presence of native MAP-2c within dendrites. In additio n, immunoelectron microscopy showed MAP-2c associated with microtubule s in dendrites of motor neurons. MAP-2c and the neurofilament proteins were found in axons of the dorsal and ventral roots. The presence of MAP-2c within axons and dendrites suggests that MAP-2c contributes to neuronal plasticity during human fetal development.